Acute diffuse axonal injury following repeated mild traumatic brain injury in juvenile rats

Mild traumatic brain injuries (mTBIs) are caused by biomechanical forces being transmitted to the brain, causing neuronal connections to be subjected to sheering forces. The injury severity can be affected by a number of factors that include age and sex, however, there remains a paucity of data on how repeated mTBI (r-mTBI) impacts the female brain. In these studies, male and female juvenile rats [<i>postnatal day</i> (PND) <i>25</i>-<i>26</i>] were administered a total of eight mTBIs over a 2-day period. Following each mTBI, rats were immediately assessed for acute neurological impairment. After eight mTBIs were completed, the Barnes maze was used to assess spatial learning and memory. Axonal injury was assessed using silver stain histological analyses. We found that injured females exhibited less acute neurological impairment than males. Three days after the final r-mTBI, no significant differences were observed in spatial learning and memory, with all animals showing similar times to locate the escape platform on the reversal trial, additionally there was no main effect of sex in the Barnes maze. Silver stain uptake was significantly increased in the optic tract, corpus callosum, and cortex compared with sham animals at seven days postinjury in a sex-specific manner. Females showed significant increase in all three regions following r-mTBI, whereas males only showed a significant increase in staining in the optic tract. Overall, these findings show that females may be more susceptible to axonal damage than males, and that cognitive deficits were not evident in this population following r-mTBI. These results indicate that there may be benefits in examining biomarkers that reflect axonal injury and the therapies that target reducing axonal degradation.<b>NEW & NOTEWORTHY</b> Diffuse axonal injury is a hallmark feature of all severities of traumatic brain injury (TBI) yet, in preclinical mild (m)TBI research no studies have yet investigated axonal damage with silver stain immunohistochemistry in female animals. This is a critical gap in the literature as recent studies suggest that females experience mTBI more frequently than males. We found that repeated mTBI (r-mTBI) caused significant diffuse axonal injury that was more pronounced in females compared with males.